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Blood, Vol. 113, Issue 23, 5776-5782, June 4, 2009
A genetic screen in zebrafish defines a hierarchical network of pathways required for hematopoietic stem cell emergence
Blood Burns et al.
113: 5776
Supplemental materials for: Burns et al
Files in this Data Supplement:
- Table S1. Four worksheets (see bottom tabs) are presented that contain the Hopkin’s Insertional line number, and whether it was identified as an HSC mutant (Mutants), ruled out (Not a Mutant), or unable to be evaluated (Unable to Evaluate) (XLS, 66 KB)
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The last worksheet shows our results from screening those mutants harboring insertions in a ribosomal protein subunit (RP Mutants). In summary, our screen identified 9 mutants that had defects in formation of c-myb+ AGM HSCs (Mutants Worksheet). We were able to rule out 185 Hopkins insertional mutant lines by whole mount in situ hybridization and/or by genotyping as described in the methods section (Not a Mutant Worksheet). There were 112 lines that were screened, but gave unclear in situ and genotyping results due to the contamination from the bloodless-like mutation (Unable to Evaluate). Of the 26 RP mutants screened, one was found to lack definitive c-myb+ HSCs, 15 had normal c-myb+ HSCs, and 10 gave unclear results and were unable to be evaluated.
- Figure S1. Characterization of definitive hematopoiesis in the mutant lines (JPG, 166 KB)
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Whole-mount in situ hybridization for c-myb, runx1, and rag-1 transcripts in 5 of the mutant lines isolated in the screen. (Column 1 and 2): Lateral views, anterior left; 36-40 hpf; Low magnification (10×) and higher magnification (40×) of the trunk region. HSC gene expression in the AGM is indicated by black arrowheads in a and b. (Column 3): Dorsal view, anterior up; 4 dpf; Bilateral rag-1 expression in thymi is indicated by black arrowheads in c.
- Figure S2. Characterization of vessel integrity and artery identity in the mutant lines (JPG, 169 KB)
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Whole-mount in situ hybridization for flk1 and efrinb2a transcripts in wild-type and mutant animals. (a–l) Lateral views, anterior left; 24–28 hpf; Low magnification (10×) and higher magnification (40×) of the trunk region. (Column 1): flk1 gene expression in the dorsal aorta is indicated by black arrowheads and intersomitic vessels are marked with green arrowheads. (Column 2): efrinb2a transcripts are highlighted with red arrowheads.
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